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  • Title: Development of an indirect enzyme immunoassay using monoclonal antibodies for the measurement of 17alpha-hydroxyprogesterone in human serum.
    Author: Chong H, Cheah SH, Ragavan M, Johgalingam VT.
    Journal: J Immunoassay Immunochem; 2009; 30(2):166-79. PubMed ID: 19330642.
    Abstract:
    An indirect enzyme immunoassay for the measurement of total 17alpha-hydroxyprogesterone (17OHP) in serum using monoclonal antibodies generated in our laboratory was developed. Here, (a) instead of extraction with solvents, serum was heated to free protein-bound 17OHP and assay was performed at pH 9.6, (b) to ensure uniform assay conditions for both standards and samples, buffer for standards contained charcoal-stripped pre-heated pooled cord serum. Assays were done in 96-well EIA microplates pre-coated with 17alpha-hydroxyprogesterone-3-(o-carboxymethyl)oxime: bovine serum albumin. Secondary antibody was horseradish peroxidase-linked sheep anti-mouse IgG polyclonal antibody. The method was accurate and suitable for screening for congenital adrenal hyperplasia.
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